PMID: 2115164May 1, 1990Paper

Structural definition by antibody engineering of an idiotypic determinant

Protein Engineering
M SollazzoM Zanetti

Abstract

Using computer-aided techniques for predicting molecular structure, we constructed an atomic model of the variable domain of a murine anti-thyroglobulin antibody whose immunodominant idiotypic determinant (Id62) was mapped by site-directed mutagenesis and immunochemical analysis. We previously showed that under experimental conditions this idiotype activates anti-idiotypic B cells and T cells, and modulates the response to thyroglobulin in mice. Because idiotype interactions are considered of physiological importance for immune regulation, we studied this idiotype as a model to understand the relationship between function and structure. To determine the contribution of heavy- and light-chain variable domains to the idiotype structure, we constructed chimeric expression vectors and introduced them into the (non-secreting) P3X63Ag8.653 myeloma cell line. Mutants of the heavy-chain variable domain were obtained by site-directed mutagenesis and transfected into the murine (lambda 1) light-chain producer J558L cell line. The expressed proteins were purified from culture supernatants of transfected cells and characterized. We provide evidence that the third hypervariable loop (D region) of the heavy-chain variable domain is the struc...Continue Reading

Citations

Jun 1, 1991·Proceedings of the National Academy of Sciences of the United States of America·R BillettaM Zanetti
Dec 15, 1993·Proceedings of the National Academy of Sciences of the United States of America·P LanzaM Zanetti
Nov 15, 2011·Nucleic Acids Research·Anna ChailyanPaolo Marcatili
Dec 1, 1992·Immunological Reviews·M ZanettiR Billetta
Sep 1, 1993·The Journal of Clinical Investigation·D Vazquez-AbadN F Rothfield
Jun 10, 1997·Proceedings of the National Academy of Sciences of the United States of America·S XiongM Zanetti
Jan 1, 1993·International Reviews of Immunology·R Billetta, M Zanetti
Dec 19, 2003·Gene Therapy·G FilaciM Zanetti
Nov 8, 2005·The Journal of Immunology : Official Journal of the American Association of Immunologists·Mara GerloniMaurizio Zanetti
Jun 21, 2008·The Journal of Immunology : Official Journal of the American Association of Immunologists·Matthew C WheelerMaurizio Zanetti
Mar 1, 2000·Methods : a Companion to Methods in Enzymology·V MoreaA Tramontano

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